RESUMEN
Objectives We investigated the effects of CD100 on naïve CD8+ T cells during hepatitis C virus (HCV) infection after interferon-α (IFN-α) therapy to clarify the mechanism underlying the effect of IFN-α in enhancing the antiviral response. Methods The CD100 molecules on subsets of CD8+ T cells were analysed with flow cytometry. The effects of CD100-overexpressing naïve CD8+ T cells were determined with ELISAs and an MTT cytotoxicity assay. The role of CD100-CD72 signal transduction was analysed with a neutralization and transwell assays. Results HCV infection reduced CD100 expression on CD8+ T cells, whereas IFN-α treatment significantly increased CD100 expression on naïve CD8+ T cells. The increased CD100 interacted with the CD72 receptor and enhanced PBMC cytokine secretion (IFN-γ and tumour necrosis factor-α) and cytotoxicity. Conclusions IFN-α-induced CD100 on naïve CD8+ T cells promotes PBMC cytokine secretion and cytotoxicity through CD100-CD72 signalling during HCV infection.
Asunto(s)
Antígenos CD/genética , Antígenos de Diferenciación de Linfocitos B/genética , Antivirales/uso terapéutico , Linfocitos T CD8-positivos/efectos de los fármacos , Hepatitis C/tratamiento farmacológico , Interferón-alfa/uso terapéutico , Polietilenglicoles/uso terapéutico , Semaforinas/genética , Adulto , Antígenos CD/inmunología , Antígenos de Diferenciación de Linfocitos B/inmunología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/virología , Estudios de Casos y Controles , Técnicas de Cocultivo , Citotoxicidad Inmunológica , Femenino , Regulación de la Expresión Génica , Hepacivirus/efectos de los fármacos , Hepacivirus/crecimiento & desarrollo , Hepacivirus/inmunología , Hepatitis C/inmunología , Hepatitis C/virología , Humanos , Interferón gamma/genética , Interferón gamma/inmunología , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/inmunología , Leucocitos Mononucleares/virología , Masculino , Persona de Mediana Edad , Cultivo Primario de Células , ARN Viral/efectos de los fármacos , Proteínas Recombinantes/uso terapéutico , Semaforinas/inmunología , Transducción de Señal , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
OBJECTIVE: To identify a novel HLA allele DRB1 * 16:36 from a Uygur woman. METHODS: PCR-SBT technology was applied to the extracted DNA for genotyping, and a possible new gene was sequenced by using sequence specific primers and single stranded SBT. This novel allele was compared with known most homologous gene sequences and their difference was analyzed. RESULTS: This novel allele was different from HLA alle DRB1 * 16:23, and had highest similarity in 2 nucleotides at position 227 AâT and 236 TâC in exon 2, resulting in 3 amino acid changes from Tyr to Phe at codon 47 and Val to Ala at codon 50. The sequence of this novel allele had been submitted to GenBank. CONCLUSION: This HLA allele DRB1 * 16:23 has been confirmed to be a novel allele, and has been officially named DRB1 * 16:36 by the World Health Organization (WHO) Nomenclature Committee in May 2015.
Asunto(s)
Alelos , Cadenas HLA-DRB1/genética , Secuencia de Bases , Cartilla de ADN , Exones , Femenino , Genotipo , Humanos , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: Borreliosis is highly prevalent in Xinjiang Uygur Autonomous Region, China. However, little is known about the presence of Borrelia pathogens in tick species in this region, in addition Borrelia pathogens have not been isolated from domestic animals. METHODS: We collected adult ticks from domestic animals at 19 sampling sites in 14 counties in northern Xinjiang from 2012 to 2014. Ticks were identified to species by morphology and were molecularly analysed by sequences of mitochondrial 16S rDNA gene; 4-8 ticks of each species at every sampling site were sequenced. 112 live adult ticks were selected for each species in every county, and were used to culture Borrelia pathogens; the genotypes were then determined by sequences of the 5S-23S rRNA intergenic spacer and the outer surface protein A (ospA) gene. RESULTS: A total of 5257 adult ticks, belonging to four genera and seven species, were collected. Compared with three decades ago, the abundance of the five common tick species during the peak ixodid tick season has changed. Certain tick species, such as Rhipicephalus turanicus (Rh. turanicus), was found at Jimusaer, Yining, Fukang, and Chabuchaer Counties for the first time. Additionally, the sequence analyses showed that the Hyalomma asiaticum (Hy. asiaticum), Haemaphysalis punctata (Ha. punctata), and Dermacentor marginatus (D. marginatus) that were collected from different sampling sites (≥3 sites) shared identical 16S rDNA sequences respectively. For the tick species that were collected from the same county, such as Hy. asiaticum from Shihezi County and Rh. turanicus from Yining County, their 16S rDNA sequences showed genetic diversity. In addition, sixteen Borrelia isolates were found in Hy. asiaticum, Ha. punctata, D. marginatus and Rh. turanicus, which infested cattle, sheep, horse and camel in Yining, Chabuchaer, Shihezi and Shawan Counties. All of the isolates were genetically identified as B. Burgdorferi sensu stricto. CONCLUSIONS: Warmer and wetter climate may have contributed to the altered distribution and abundance of the five most common ticks in northern Xinjiang. The genetic analyses showed that certain tick species, such as Hy. asiaticum or Rh. turanicus, exhibit genetic commonness or diversity. Additionally, this study is the first to isolate B. burgdorferi sensu stricto in Hy. asiaticum asiaticum, H. punctata, D. nuttalli and D. marginatus ticks from domestic animals. These ticks may transmit borreliosis among livestock.
Asunto(s)
Borrelia burgdorferi/aislamiento & purificación , Ganado/parasitología , Infestaciones por Garrapatas/veterinaria , Garrapatas/clasificación , Garrapatas/microbiología , Animales , China/epidemiología , Filogenia , Especificidad de la Especie , Infestaciones por Garrapatas/epidemiología , Infestaciones por Garrapatas/parasitologíaRESUMEN
Beijing/W lineage strains of Mycobacterium tuberculosis spread faster than other strains, tend to be more virulent and frequently associated with drug resistance. In this study, to distinguish the characteristics of Beijing/W lineage and non-Beijing/W lineage M. tuberculosis, we assessed the growth between the two groups under conditions of hypoxia, nutrient starvation, and intracellular growth in murine macrophages. We also examined the DNA, RNA, and protein levels of 5 major M. tuberculosis proteins, including HspX, Hsp65, 38 kDa, Ag85B, and MPT64 of the different types of strains by sequencing, quantitative RT-PCR, and Western blotting. The results showed that Beijing/W and non-Beijing/W lineage strains of M. tuberculosis have similar viability in ex vivo culture but differ in their ability to survive within macrophages, and the intracellular viability of the Beijing/W lineage strains was significantly more than the viability of the non-Beijing/W lineage strains at 2, 3, and 5 days after infection (P < 0.05). Psts1 and fbpB were expressed at statistically lower levels in Beijing/W lineage strains in their mRNA expression levels (P < 0.05). The expression of their corresponding 38 kDa and Ag85B was lower in the Beijing/W lineage strains than the non-Beijing/W lineage strains (P < 0.05). The expression of HspX and Hsp65 was higher in the Beijing/W lineage strains in their protein expression levels at 24 h after infection of RAW264.7 macrophages (P < 0.05). In conclusion, the increased viability of the Beijing/W lineage strains might be related to the expression levels of these proteins.